Experiment
JoVE Encyclopedia of Experiments
Encyclopedia of Experiments: Biological Techniques
Encyclopedia of Experiments: Biological Techniques
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Anion-Exchange Chromatography-Based Protein Purification
Transcript
Start the FPLC with general maintenance. Connect column buffers AIEX-A and AIEX-B to buffer valves, and the anion-exchange resin-containing column to the column ports of the FPLC. Adjust the general chromatographic parameters accordingly.
Start the AIEX program in the FPLC software and choose the equilibration volume for buffer A. Subsequently, load the sample onto the column, and elute the proteins with a linear gradient from 0% to 100% high-salt buffer. Collect 2-milliliter fractions during elution.
Analyze the eluted fractions after finishing the run.
